P
US7507718B2ExpiredUtilityPatentIndex 43

Polymyxin B analogs for LPS detoxification

Assignee: SANOFI PASTEURPriority: Apr 11, 2005Filed: Apr 6, 2006Granted: Mar 24, 2009
Est. expiryApr 11, 2025(expired)· nominal 20-yr term from priority
Inventors:PORRO MASSIMOVELUCCHI MASSIMORUSTICI ALESSANDROMOREAU MONIQUEMISTRETTA NOELLEKRELL TINO
Y02A50/30C07K 7/54C07K 7/06A61K 38/00C07K 7/08
43
PatentIndex Score
1
Cited by
21
References
77
Claims

Abstract

The invention relates to SAEP II peptide dimers that mimic polymyxin B i.a. in its ability to bind non-covalently the lipopolysaccharide (LPS) of Gram-negative bacteria with high affinity, and therefore to detoxify LPS. The dimeric structure is maintained by a pair of disulphide bonds between two cystein residues present in the peptide sequence, which does not exceed 17 amino acids and essentially comprises cationic and hydrophobic amino acid residues. The peptides in the dimers may have a parallel or anti-parallel orientation. SAEP II dimers are useful for treating or preventing septic shock and related disorders generated by Gram-negative bacteria infection. The invention also relates to LPS-peptide complexes in which LPS and SAEP II diners are non-covalently bound together. These complexes are useful as vaccinal agents against Gram-negative bacteria infection.

Claims

exact text as granted — not AI-modified
1. A SAEP II peptide dimer, wherein the peptide dimer is essentially in the form of
 (a) a dimer of formula (I)
   NH 2 -A-Cys1-B-Cys2-C—COOH 
   NH 2 -A′-Cys1-B′-Cys2-C′—COOH 
 
 wherein Cys1 and Cys2 are each a cysteine amino acid; 
 wherein the two Cys1 residues are linked together through a disulphide bond and the two Cys2 residues are linked together through a disulphide bond; 
 and/or 
 (b) a dimer of formula (II)
   NH 2 -A-Cys1-B-Cys2-C—COOH 
   HOOC—C′-Cys2-B′-Cys1-A′-NH 2   
 
 wherein Cys1 and Cys2 are each a cysteine amino acid; 
 wherein the Cys1 residues are linked to the Cys2 residues through intermolecular disulphide bonds; 
 wherein A and A′ independently are a peptide moiety of from 2 to 5 amino acid residues, in which at least 2 amino acid residues are independently selected from Lys, Hyl (hydroxy-Lysine), Arg and His; 
 wherein B and B′ independently are a peptide moiety of from 3 to 7 amino acid residues, which comprise at least two amino acid residues independently selected from Val, Leu, Ile, Phe, Tyr and Trp; and 
 wherein C and C′ are optional and are independently an amino acid residue or a peptide moiety of from 2 to 3 amino acid residues; 
 provided that the cationic amino acid residues/hydrophobic amino acid residues ratio (cat/hydroph ratio) of the dimer is from 0.4 to 2. 
 
     
     
       2. The SAEP II peptide dimer according to  claim 1 , wherein the cat/hydroph ratio is from 0.5 to 1.5. 
     
     
       3. The SAEP II peptide dimer according to  claim 2 , wherein the cat/hydroph ratio is from 0.6 to 1. 
     
     
       4. The SAEP II peptide dimer according to  claim 3 , wherein the cat/hydroph ratio is from 0.6 to 0.8. 
     
     
       5. The SAEP II peptide dimer according to  claim 1 , wherein the B and B′ peptide moieties comprise the sequence -X1-X2-X3-, in which X1 and X2; X2 and X3; or X1, X2 and X3 are independently selected from Val, Leu, Ile, Phe, Tyr and Trp. 
     
     
       6. The SAEP II peptide dimer according to  claim 5 , wherein the B and B′ peptide moieties independently comprise:
 (i) the sequence -X1-X2-X3-, in which: 
 X1 is Lys, Hyl, His or Arg; 
 X2 is Phe, Leu, Ile, Tyr, Trp or Val; and 
 X3 is Phe, Leu, Ile, Tyr, Trp or Val; and 
 (ii) if the B and B′ peptide moieties comprise amino acid residues, in addition to the sequence -X1-X2-X3-, the additional amino acid residues are independently selected from the group consisting of Val, Leu, Ile, Phe, Tyr, Trp, Lys, Hyl, Arg and His. 
 
     
     
       7. The SAEP II peptide dimer according to  claim 1 , which is a dimer of formula (III)
   NH 2 -A-Cys1-B-Cys2-COOH 
   NH 2 -A′ -Cys1-B′-Cys2-COOH 
 wherein the two Cys1 residues are linked together through a disulphide bond and the two Cys2 residues are linked together through a disulphide bond; 
 and/or formula (IV)
   NH 2 -A-Cys1-B-Cys2-COOH 
   HOOC-Cys2-B′-Cys1-A′-NH 2 , 
 
 wherein the Cys1 residues are linked to the Cys2 residues through intermolecular disulphide bonds. 
 
     
     
       8. The SAEP II peptide dimer according to  claim 1 , which is a homologous peptide dimer. 
     
     
       9. The SAEP II peptide dimer according to  claim 1 , which is an antiparallel dimer form of formula (VI)
   NH 2 -Lys-Thr-Lys-Cys1-Lys-Phe-Leu-Leu-Leu-Cys2-COOH 
   COOH-Cys2-Leu-Leu-Leu-Phe-Lys-Cys1-Lys-Thr-Lys-NH 2 , 
 wherein the Cys1 residues are linked to the Cys2 residues through intermolecular disulphide bonds. 
 
     
     
       10. The SAEP II peptide dimer according to  claim 1 , which is a parallel dimer of formula (VII)
   NH 2 -Lys-Thr-Lys-Cys1-Lys-Phe-Leu-Leu-Leu-Cys2-COOH 
   NH 2 -Lys-Thr-Lys-Cys1-Lys-Phe-Leu-Leu-Leu-Cys2-COOH 
 wherein the two Cys1 residues are linked together through a disulphide bond and the two Cys2 residues are linked together through a disulphide bond. 
 
     
     
       11. A composition comprising a peptide dimer according to  claim 1 , wherein the peptide dimer is essentially in dimeric parallel form. 
     
     
       12. A composition comprising a peptide dimer according to  claim 1 , wherein the peptide dimer is essentially in dimeric antiparallel form. 
     
     
       13. A pharmaceutical composition comprising (i) a peptide dimer according to  claim 1  and (ii) a pharmaceutically acceptable diluent or carrier. 
     
     
       14. A method for treating or preventing septic shock, which comprises administering a therapeutically effective amount of a peptide dimer according to  claim 1  to an individual in need. 
     
     
       15. A LPS-peptide complex comprising (i) a LPS entity of Gram-negative bacteria and (ii) a SAEP II peptide dimer according to  claim 1 , wherein the LPS moiety and the SAEP II peptide are non-covalently bound to each other. 
     
     
       16. The LPS-peptide complex according to  claim 15 , in which the LPS is a LPS of  N. meningitidis; E. coli; Salmonella typhi; Salmonella paratyphi; Shigella fexneri; Haemophilus influenzae, Helicobacter pylori; Chlamydia trachomatis; Bordetella pertussis; Brucella; Legionella pneumophia; Vibrio cholera; Moraxella catharralis; Pseudomonas aeruginosa;  and  Kiebsiella pneumonia.    
     
     
       17. The LPS-peptide complex according to  claim 16 , in which the LPS is a LPS of  Neisseria meningitidis.    
     
     
       18. The LPS-peptide complex according to  claim 17 , in which the LPS is LPS L8. 
     
     
       19. The LPS-peptide complex according to  claim 17 , characterized by a molar LPS:peptide ratio of from 1:1.5 to 1:0.5. 
     
     
       20. The LPS-peptide complex according to  claim 19 , characterized by a molar LPS:peptide ratio of 1:1. 
     
     
       21. A pharmaceutical composition comprising a LPS-peptide complex according to  claim 15 , and a pharmaceutically acceptable diluent or carrier. 
     
     
       22. A method for treating or preventing a Gram-negative bacterial infection, which comprises administering a therapeutically effective amount of a LPS-peptide complex according to  claim 15 , to an individual in need thereof. 
     
     
       23. A process for preparing a LPS/peptide complex, which comprises mixing together (i) a LPS of Gram-negative bacteria and (ii) a peptide dimer according to  claim 1 . 
     
     
       24. The process according to  claim 23 , wherein the LPS and the peptide dimer or salt thereof are mixed in a molar LPS:peptide ratio of from 1:1.2 to 1:0.8. 
     
     
       25. The process according to  claim 24 , wherein the LPS and the peptide dimer or salt thereof are mixed in a 1:1 molar LPS:peptide ratio. 
     
     
       26. A process for detoxifying a LPS of Gram-negative bacteria, which comprises mixing together (i) the LPS and (ii) a peptide dimer according to  claim 1 . 
     
     
       27. The process according to  claim 26 , wherein the LPS and the peptide dimer are mixed in a molar LPS:peptide ratio of from 1:1.2 to 1:0.8. 
     
     
       28. The process according to  claim 27 , wherein the LPS and the peptide dimer or salt thereof are mixed in a 1:1 molar LPS:peptide ratio. 
     
     
       29. The SAEP II peptide dimer according to  claim 1 , wherein the dimer is purified from non-dimer peptides. 
     
     
       30. The SAEP II peptide dimer according to  claim 1 , wherein B and B′ comprise at least three amino acid residues independently selected from Val, Leu, Ile, Phe, Tyr and Trp. 
     
     
       31. The SAEP II peptide dimer according to  claim 30 , wherein the dimer is purified from non-dimer peptides. 
     
     
       32. A SAEP II peptide dimer, wherein the peptide dimer is at least 95% in the form of
 (a) a dimer of formula (I)
   NH 2 -A-Cys1-B-Cys2-C-COOH 
   NH 2 -A′-Cys1-B′-Cys2-C′-COOH 
 
 wherein Cys1 and Cys2 are each a cysteine amino acid; 
 wherein the two Cys1 residues are linked together through a disulphide bond and the two Cys2 residues are liniked together through a disulphide bond; 
 and/or 
 (b) a dimer of formula (II)
   NH 2 -A-Cys1-B-Cys2-C-COOH 
   HOOC-C′-Cys2-B′-Cys1-A′-NH 2   
 
 wherein Cys1 and Cys2 are each a cysteine amino acid; 
 wherein the Cys1 residues are linked to the Cys2 residues through intermolecular disulphide bonds; 
 wherein A and A′ independently are a peptide moiety of from 2 to 5 amino acid residues, in which at least 2 amino acid residues are independently selected from Lys, Hyl (hydroxy-Lysine), Arg and His; 
 wherein B and B′ independently are a peptide moiety of from 3 to 7 amino acid residues, which comprise at least two amino acid residues independently selected from Val, Leu, Ile, Phe, Tyr and Trp; and 
 wherein C and C′ are optional and are independently an amino acid residue or a peptide moiety of from 2 to 3 amino acid residues; 
 provided that the cationic amino acid residues/hydrophobic amino acid residues ratio (cat/hydroph ratio) of the dimer is from 0.4 to 2. 
 
     
     
       33. The SAEP II peptide dimer according to  claim 32 , wherein the cat/hydroph ratio is from 0.5 to 1.5. 
     
     
       34. The SAEP II peptide dimer according to  claim 33 , wherein the cat/hydroph ratio is from 0.6 to 1. 
     
     
       35. The SAEP II peptide dimer according to  claim 34 , wherein the cat/hydroph ratio is from 0.6 to 0.8. 
     
     
       36. The SAEP II peptide dimer according to  claim 32 , wherein the B and B′ peptide moieties comprise the sequence -X1-X2-X3-, in which X1 and X2; X2 and X3; or X1, X2 and X3 are independently selected from Val, Leu, Ile, Phe, Tyr and Trp. 
     
     
       37. The SAEP II peptide dimer according to  claim 36 , wherein the B and B′ peptide moieties comprise:
 (i) the sequence -X1-X2-X3-, in which: 
 X1 is Lys, Hyl, His or Arg; 
 X2 is Phe, Leu, Ile, Tyr, Trp or Val; and 
 X3 is Phe, Leu, Ile, Tyr, Trp or Val; and 
 (ii) if the B and B′ peptide moieties comprise amino acid residues, in addition to the seciuence -X1-X2-X3-, the additional amino acid residues are independently selected from the group consisting of Val, Leu, Ile, Phe, Tyr, Trp, Lys, Hyl, Arg and His. 
 
     
     
       38. The SAEP II peptide dimer according to  claim 32 , which is a dimer of formula (III)
   NH 2 -A-Cys1-B-Cys2-COOH 
   NH 2 -A′-Cys1-B′-Cys2-COOH 
 wherein the two Cys1 residues are linked together through a disulphide bond and the two Cys2 residues are linked together through a disulphide bond; and/or formula (IV)
   NH 2 -A-Cys1-B-Cys2-COOH 
   HOOC-Cys2-B′-Cys1-A′-NH 2 , 
 
 wherein the Cys1 residues are linked to the Cys2 residues through intermolecular disulphide bonds. 
 
     
     
       39. The SAEP II peptide dimer according to  claim 32 , which is a homologous peptide dimer. 
     
     
       40. The SAEP II peptide dimer according to  claim 32 , which is an antiparallel dimer form of formula (VI)
   NH 2 -Lys-Thr-Lys-Cys1-Lys-Phe-Leu-Leu-Leu-Cys2-COOH 
   COOH-Cys2-Leu-Leu-Leu-Phe-Lys-Cys1-Lys -Thr-Lys-NH 2 , 
 wherein the Cys1 residues are linked to the Cys2 residues through intermolecular disulphide bonds. 
 
     
     
       41. The SAEP II peptide dimer according to  claim 32 , which is a parallel dimer of formula (VII)
   NH 2 -Lys-Thr-Lys-Cys1-Lys-Phe-Leu-Leu-Leu-Cys2-COOH 
   NH 2 -Lys-Thr-Lys-Cys1-Lys-Phe-Leu-Leu-Leu-Cys2-COOH 
 wherein the two Cys1 residues are linked together through a disulphide bond and the two Cys2 residues are linked together through a disulphide bond. 
 
     
     
       42. A composition comprising a peptide dimer according to  claim 32 , wherein the peptide is essentially in dimeric parallel form. 
     
     
       43. A composition comprising a peptide dimer according to  claim 32 , wherein the peptide is essentially in dimeric antiparallel form. 
     
     
       44. A pharmaceutical composition comprising (i) a peptide dimer according to  claim 32  and (ii) a pharmaceutically acceptable diluent or carrier. 
     
     
       45. A SAEP II peptide dimer, wherein the peptide is essentially in the form of
 (a) a dimer of-formula (I)
   NH 2 -A-Cys1-B-Cys2-C-COOH 
   NH 2 -A′-Cys1-B′-Cys2-C′-COOH 
 
 wherein Cys1 and Cys2 are each a cysteine amino acid; 
 wherein the two Cys1 residues are linked together through a disulphide bond and the two Cys2 residues are linked together through a disulphide bond; 
 and/or 
 (b) a dimer of formula (II)
   NH 2 -A-Cys1-B-Cys2-C-COOH 
   HOOC-C′-Cys2-B′-Cys1-A′-NH 2   
 
 wherein Cys1 and Cys2 are each a cysteine amino acid; 
 wherein the Cys1 residues are linked to the Cys2 residues through intermolecular disulphide bonds; 
 wherein A and A′ independently are a peptide moiety of from 2 to 5 amino acid residues, in which at least 2 amino acid residues are independently selected from Lys, Hyl (hydroxy-Lysine), Arg and His; 
 wherein B and B′ independently are a peptide moiety of from 3 to 7 amino acid residues, which comprise at least two amino acid residues independently selected from Val, Leu, Ile, Phe, Tyr and Trp; and 
 wherein C and C′ are optional and are independently an amino acid residue or a peptide moiety of from 2 to 3 amino acid residues; 
 provided that the cationic amino acid residues/hydrophobic amino acid residues ratio (cat/hydroph ratio) of the dimer is from 0.4 to 2, and 
 wherein the dimer is in solid form. 
 
     
     
       46. A pharmaceutical composition comprising (i) a peptide dimer according to  claim 45  and (ii) a pharmaceutically acceptable diluent or carrier. 
     
     
       47. The SAEP II peptide dimer according to any one of  claims 1  or  32 , which is a heterodimer. 
     
     
       48. A LPS-peptide complex comprising (i) a LPS entity (moiety) of Gram-negative bacteria and (ii) a SAEP II peptide dimer according to  claim 32 , wherein the LPS moiety and the SAEP II peptide are non-covalently bound to each other. 
     
     
       49. The LPS-peptide complex according to  claim 48 , in which the LPS is a LPS of  N. meningitidis; E. coli; Salmonella typhi; Salmonella paratyphi; Shigella fexneri; Haemophilus influenzae; Helicobacter pylori; Chlamydia trachomatis; Bordetella pertussis; Brucella; Legionella pneumophia; Vibrio cholera; Moraxella catharralis; Pseudomonas aeruginos;  and  Kiebsiella pneumonia.    
     
     
       50. The LPS-peptide complex according to  claim 49 , in which the LPS is a LPS of  Neisseria meningitidis.    
     
     
       51. The LPS-peptide complex according to  claim 50 , in which the LPS is LPS L8. 
     
     
       52. The LPS-peptide complex according to  claim 50 , characterized by a molar LPS:peptide ratio of from 1:1.5 to 1:0.5. 
     
     
       53. The LPS-peptide complex according to  claim 52 , characterized by a molar LPS:peptide ratio of 1:1. 
     
     
       54. A pharmaceutical composition comprising a LPS-peptide complex according to  claim 48 , and a pharmaceutically acceptable diluent or carrier. 
     
     
       55. A process for preparing a LPS/peptide complex, which comprises mixing together (i) a LPS of Gram-negative bacteria and (ii) a peptide dimer according to  claim 32 . 
     
     
       56. The process according to  claim 55 , wherein the LPS and the peptide dimer or salt thereof are mixed in a molar LPS:peptide ratio of from 1:1.2 to 1:0.8. 
     
     
       57. The process according to  claim 56 , wherein the LPS and the peptide dimer or salt thereof are mixed in a 1:1 molar LPS:peptide ratio. 
     
     
       58. A LPS-peptide complex comprising (i) a LPS entity (moiety) of Gram-negative bacteria and (ii) a SAEP II peptide dimer according to  claim 9 , wherein the LPS moiety and the SAEP II peptide are non-covalently bound to each other. 
     
     
       59. The LPS-peptide complex according to  claim 58 , in which the LPS is a LPS of  N. meningitidis; E. coli; Salmonella typhi; Salmonella paratyphi; Shingella fexneri; Haemophilus influenzae; Helicobacter pylori; Chlamydia trachomatis; Bordetella pertussis; Brucella; Legionella pneumophia; Vibrio cholera; Moraxella catharralis; Pseudomonas aeruginosa;  and  Kiebsiella pneumonia.    
     
     
       60. The LPS-peptide complex according to  claim 59 , in which the LPS is a LPS of  Neisseria meningitidis.    
     
     
       61. The LPS-peptide complex according to  claim 60 , in which the LPS is LPS L8. 
     
     
       62. The LPS-peptide complex according to  claim 60 , characterized by a molar LPS:peptide ratio of from 1:1.5 to 1:0.5. 
     
     
       63. The LPS-peptide complex according to  claim 62 , characterized by a molar LPS:peptide ratio of 1:1. 
     
     
       64. A pharmaceutical composition comprising a LPS-peptide complex according to  claim 58 , and a pharmaceutically acceptable diluent or carrier. 
     
     
       65. A process for preparing a LPS/peptide complex, which comprises mixing together (i) a LPS of Gram-negative bacteria and (ii) a peptide dimer according to  claim 9 . 
     
     
       66. The process according to  claim 65 , wherein the LPS and the peptide dimer or salt thereof are mixed in a molar LPS:peptide ratio of from 1:1.2 to 1:0.8. 
     
     
       67. The process according to  claim 66 , wherein the LPS and the peptide dimer or salt thereof are mixed in a 1:1 molar LPS:peptide ratio. 
     
     
       68. A LPS-peptide complex comprising (i) a LPS entity (moiety) of Gram-negative bacteria and (ii) a SAEP II peptide according to  claim 40 , wherein the LPS moiety and the SAEP II peptide dimer are non-covalently bound to each other. 
     
     
       69. The LPS-peptide complex according to  claim 68 , in which the LPS is a LPS of  N. meningitidis; E. coli; Salmonella typhi; Salmonella paratyphi; Shingella fexneri; Haemophilus influenzae; Helicobacter pylori; Chlamydia trachomatis; Bordetella pertussis; Brucella; Legionella pneumophia, Vibrio cholera, Moraxella catharralis, Pseudomonas aeruginosa,  and  Kiebsiella pneumonia.    
     
     
       70. The LPS-peptide complex according to  claim 69 , in which the LPS is a LPS of  Neisseria meningitidis.    
     
     
       71. The LPS-peptide complex according to  claim 70 , in which the LPS is LPS L8. 
     
     
       72. The LPS-peptide complex according to  claim 70 , characterized by a molar LPS:peptide ratio of from 1:1.5 to 1:0.5. 
     
     
       73. The LPS-peptide complex according to  claim 72 , characterized by a molar LPS:peptide ratio of 1:1. 
     
     
       74. A pharmaceutical composition comprising a LPS-peptide complex according to  claim 68 , and a pharmaceutically acceptable diluent or carrier. 
     
     
       75. A process for preparing a LPS/peptide complex, which comprises mixing together (i) a LPS of Gram-negative bacteria and (ii) a peptide dimer according to  claim 40 . 
     
     
       76. The process according to  claim 75 , wherein the LPS and the peptide dimer or salt thereof are mixed in a molar LPS:peptide ratio of from 1:1.2 to 1:0.8. 
     
     
       77. The process according to  claim 76 , wherein the LPS and the peptide dimer or salt thereof are mixed in a 1:1 molar LPS:peptide ratio.

Cited by (0)

No later patents cite this yet.

References (0)

No backward citations on record.